Team:Wisconsin-Madison

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    <p class = "classtheoverview"> Engineering the limonene production metabolic pathway: TCC. </p>
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      <p class = "classtheoverview"> <strong> The Translational Coupling Cassette: a tool for evaluating the translation of heterologous proteins in <i>Escherichia coli</i>. </strong></p>
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       <p class = "classtheoverview"> Creating a tool to evaluate the translation of heterologous genes in Eschericia coli. </p>
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       <p align="left" class = "classtheinlinecontent2"> A powerful method for the production of novel metabolites is the expression of heterologous enzymes in a bacterial host. A common challenge when using non-native genes in metabolic engineering is determining if they are being properly expressed. To address this issue, we have constructed a BioFusion-compatible system for testing the translation of a gene of interest. This system couples the translation of the target gene to a fluorescent reporter gene; fluorescence will only be detected when the target gene is entirely translated. This construct enables synthetic biologists to quickly determine if a gene is being expressed without the need for costly antibodies or analytical instruments (e.g. mass spectrometry). Currently, we are utilizing this cassette to optimize the expression of limonene synthase, an enzyme that catalyzes the production of limonene, a monoterpene with potential as a renewable jet fuel.</p>
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      <p align="left" class="classtheinlinecontent">In synthetic biology, one of the most powerful tools a research has is the ability to express heterologous genes in Escherichia coli. Unfortunately this does not always work. A common challenge when using non-native genes in metabolic engineering is determining if they are being properly expressed. To address this issue, we have constructed a BioFusion compatible system for diagnosing the failure of a gene expression at the translational level. This system couples the translation of the target gene to a fluorescent reporter gene.  Fluorescence will only be detected when the target gene is entirely translated. This construct enables synthetic biologists to quickly determine if a gene is being expressed without the need for costly antibiodies or analytical instruments.</p>
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<p align="left" class="classtheinlinecontent" style="font-style:italic">This tool is based on a phenomenon known as translational coupling. </p>
 
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Latest revision as of 20:08, 26 October 2012


The Translational Coupling Cassette: a tool for evaluating the translation of heterologous proteins in Escherichia coli.

A powerful method for the production of novel metabolites is the expression of heterologous enzymes in a bacterial host. A common challenge when using non-native genes in metabolic engineering is determining if they are being properly expressed. To address this issue, we have constructed a BioFusion-compatible system for testing the translation of a gene of interest. This system couples the translation of the target gene to a fluorescent reporter gene; fluorescence will only be detected when the target gene is entirely translated. This construct enables synthetic biologists to quickly determine if a gene is being expressed without the need for costly antibodies or analytical instruments (e.g. mass spectrometry). Currently, we are utilizing this cassette to optimize the expression of limonene synthase, an enzyme that catalyzes the production of limonene, a monoterpene with potential as a renewable jet fuel.









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